基因工程第六章 DNA Sequence.ppt

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Confidential and Proprietary—DO NOT DUPLICATE * Ion Technology /technology-chemistry-video/ Confidential and Proprietary DO NOT DUPLICATE * The Chip is the Machine? Scalability Simplicity Speed Simplicity Confidential and Proprietary—DO NOT DUPLICATE * Highly uniform genome coverage Rapidly improving raw accuracy Measured, not predicted and includes homopolymers Low-cost reagents and detection Post-Light? sequencing enables rapid improvement Simple Post-Light? Sequencing System Simple Chemistry Confidential and Proprietary—DO NOT DUPLICATE * Fast Direct Detection DNA ? Ions ? Sequence Nucleotides flow sequentially over Ion semiconductor chip One sensor per well per sequencing reaction Direct detection of natural DNA extension Millions of sequencing reactions per chip Fast cycle time, real time detection Confidential and Proprietary—DO NOT DUPLICATE * Sensor Plate Silicon Substrate Drain Source Bulk dNTP To column receiver ? pH ? Q ? V Sensing Layer H+ * * * * * * * * * * * * * * * * * * * * * * * * * * if then (Sn=50%, n10) L=8000 , ?=400 , Sn=98% n=76 if then n=101309 L=5,500,000 , ?=500, Sn=99.99% N=log(1-Sn)/log(1-?/L) coverage 9.21 times 3. Fixed cloned Constructing physical map 4. Deletion clones Erase-a-Base? System Promega Exonuclease III 5‘ 3‘ 5‘ 3‘ 5‘ 3‘ 第四节 RNA端点分析 1. S1酶作图 标记寡核苷酸, PAGE电泳检测MW 5‘ 3‘ ATG UAG S1酶 S1酶 还可检测前体RNA-mRNA的剪接点 3’AGCUCGUAGCAUGCAU5’ CATCGTA CATCGTACGTAG CATCGTAC CATCGTACG CATCGTACGT CATCGTACGTA 3’AGCTCGTAGCATGCATCGATGCATGCTAGCAT 5’ DNA RNA CATCGTACGTA 2 Primer extension T T T A A G T G A A A A A A C A G C T* T A A A T T C C A A A A G C A T T A T A G A A G T* A T C A A T T T T G C A Primer extension for CTC gene Solexa SOLiD 100MB 1GB 4G 250bp 35bp 35bp 新一代测序技术 2005年底，罗氏诊断公司和454公司推出了—